Tryptophan-130 is the most reactive tryptophan residue in rabbit skeletal myosin subfragment-1.
Rabbit skeletal muscle myosin subfragment-1 (S-1) was reacted with dimethyl(2-hydroxy-5-nitrobenzyl)sulfonium bromide (DHNBS) resulting in modification of 0.8 tryptophan residues per S-1. In order to assign the most reactive tryptophan of the 5 S-1 tryptophans, antibodies were raised in rabbits against bovine serum albumin modified with DHNBS. The antibodies reacted with the 27 kDa tryptic fragment of DHNBS-treated S-1, indicating that the reactive tryptophan resides on this domain. The 27 kDa fragment was isolated from DHNBS-treated S-1 and was further cleaved at a single cysteine residue by 2-nitro-5-thiocyanobenzoic acid. This cleavage resulted in two peptides, each of them containing one tryptophan. The antibodies reacted with the smaller peptide consisting of residues 122-204. The only tryptophan residing on this peptide is Trp130, and this is therefore the most reactive tryptophan of S-1.
Peyser
Y M
YM
Department of Oral Biology, Hebrew University-Hadassah School of Dental Medicine, Jerusalem, Israel.
Muhlrad
A
A
Werber
M M
MM
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007202:Indicators and Reagents; D008970:Molecular Weight; D009132:Muscles / Q000378:metabolism*; D015879:Myosin Subfragments / Q000378:metabolism*; D009861:Onium Compounds / Q000378:metabolism*; D011485:Protein Binding; D011817:Rabbits; D013452:Sulfonium Compounds / Q000378:metabolism*; D014357:Trypsin; D014364:Tryptophan
D007202:Indicators and Reagents; D015879:Myosin Subfragments; D009861:Onium Compounds; D013452:Sulfonium Compounds; C024586:dimethyl(2-hydroxy-5-nitrobenzyl)sulfonium; D014364:Tryptophan; D014357:Trypsin